Systems and methods for cleaning and disinfecting allograft material

ABSTRACT

There is disclosed a system for cleaning and disinfecting allograft material. In an embodiment, the system includes a sterile bag and a motorized paddle blender. The sterile bag having an outer wall configured to withstand blending forces, at least one paddle contact section, and an allograft retaining section separate from the at least one paddle contact section. The motorized paddle blender having at least one paddle configured to apply blending forces on the sterile bag on the at least one paddle contact area, a motorized portion to actuate the at least one paddle, and a door component configured to hold the sterile bag adjacent the at least one paddle. The at least one paddle and the allograft retaining section are configured to prevent the paddle from contacting the allograft material when the blending forces are applied on the sterile bag on the at least one paddle contact area.

REFERENCE TO PENDING PRIOR PATENT APPLICATION

This application is a continuation-in-part of prior U.S. patentapplication Ser. No. 13/449,636, filed Apr. 18, 2012 by Jan Zajdowiczfor SYSTEMS AND METHODS FOR CLEANING AND DISINFECTING ALLOGRAFTMATERIAL, which issued on Sep. 22, 2015 as U.S. Pat. No. 9,138,496.

BACKGROUND

The use of musculoskeletal allograft tissue in reconstructive orthopedicprocedures and other medical procedures has markedly increased over thelast decade. The most common allograft is bone. However, tendons, skin,heart valves and corneas are other common types of tissue allografts.

Prior to use, the allograft tissue must be evaluated for microbialcontamination. The allograft product must be tested for bacterialcontamination prior to release of the tissue for transplantation. Swabsare widely used in the pharmaceutical and medical device industry forevaluating microbial contaminants on small, hard, non-porousmanufacturing equipment, in addition to detecting microbial contaminantsin environmental monitoring programs. Some areas of the allograft aresimply inaccessible to a swab, thereby not allowing for completeanalysis of the allograft for microbial contaminants.

Another method used for detecting microbial contamination on allograftsis destructive testing. Destructive testing using companion tissues(small sections of typically lower quality or unusable portions of theallograft) is routinely used to assess microbial contamination on entireallograft lots. This practice has come under intense scrutiny byregulatory agencies since the companion tissue may not be representativeof the microbial contamination on entire allograft lot. Furthermore, thegeometry of the companion tissue does not adequately represent thegeometry of the entire allograft lot.

Recently, non-allograft materials from varying sources (bovine, ceramic,synthetic, etc.) have been used as a representative model of what theallograft tissue products are exposed to during handling and processing.The limitation with these materials is that they are not trulyrepresentative of the actual allograft. Furthermore, it is extremelydifficult to fabricate synthetic samples to model every product categorycurrently utilized for transplantation.

Prior to use, the allograft tissue must be treated with various agentsin order to substantially eliminate microbial contamination as well asclean the tissue of residual blood constituents, bone marrow, residualconnective tissue and gross musculature. A variety of cleaning processeshave been developed in order to remove contaminants from the allograftand to inactivate microbial contaminants remaining on the allografts.However, these cleaning and inactivation methods are laborious andtedious, and often do not provide a high level of assurance that theallografts have been sufficiently cleaned (e.g., low or inconsistent logreductions in microbial contamination). In particular, many existingallograft cleaning processes require considerable manipulation of theallografts between steps, thus increasing the possibility ofenvironmental cross-contamination. Existing processes also tend to behard to regulate and control, and their efficacy can be techniciandependent. Existing processes also tend to have a shielding or layeringeffect that can greatly reduce ultrasonic energy penetration and thusnot clean as effectively. Furthermore, the shielding effect will alsoimpede the liberation of contaminant microorganisms off of the tissuesand into solution where they are more readily eradicated.

Following treatment, allograft products must be tested for bacterialcontamination prior to release of the tissue for transplantation.Existing methods of assessing microbial contamination, however, sufferform the same limitations described above (e.g. considerablemanipulation between steps, possibility for environmentalcross-contamination, hard to regulate and control, technician dependent,etc.).

In the past, ultrasound has been utilized to reduce and/or eliminatemicrobial contamination of allograft products. Ultrasound ismicrobiostatic to most microbes, and is used primarily to reducemicrobial loads from inanimate objects with specific bactericidalactivity on gram-negative bacteria.

With the increased use of allograft products, there is a need to provideimproved methods and apparatus for treating allografts in order to helpprovide the cleanest and safest allografts as well as confirm that theallografts are free from bacterial contamination.

SUMMARY

This Summary is provided to introduce a selection of concepts in asimplified form that are further described below in the DetailedDescription. This Summary is not intended to identify key aspects oressential aspects of the claimed subject matter. Moreover, this Summaryis not intended for use as an aid in determining the scope of theclaimed subject matter.

In an embodiment, there is provided a system for cleaning anddisinfecting allograft material, the system comprising a sterile baghaving an outer wall configured to withstand blending forces, at leastone paddle contact section, and an allograft retaining section separatefrom the at least one paddle contact section; a motorized paddle blenderhaving at least one paddle configured to apply blending forces on thesterile bag on the at least one paddle contact area, a motorized portionto actuate the at least one paddle, and a door component configured tohold the sterile bag adjacent the at least one paddle, wherein the atleast one paddle and the allograft retaining section are configured toprevent the paddle from contacting the allograft material when theblending forces are applied on the sterile bag on the at least onepaddle contact area.

In another embodiment, there is provided a process of cleaning allograftmaterial, the process comprising staged, continuous, or both staged andcontinuous solution fluid pumping into the bag system.

In yet another embodiment, there is provided a process of disinfectingallograft material with an allograft microbial extraction process.

In still another embodiment, there is provided a process ofdemineralizing allograft material with an acid solution.

In another embodiment, there is provided a process of preparingallograft material with an allograft viability promotion process.

In still yet another embodiment, there is provided a process ofpreparing allograft material with a final packaging process.

Other embodiments are also disclosed.

Additional objects, advantages and novel features of the technology willbe set forth in part in the description which follows, and in part willbecome more apparent to those skilled in the art upon examination of thefollowing, or may be learned from practice of the technology.

BRIEF DESCRIPTION OF THE DRAWINGS

Non-limiting and non-exhaustive embodiments of the present invention,including the preferred embodiment, are described with reference to thefollowing figures, wherein like reference numerals refer to like partsthroughout the various views unless otherwise specified. Illustrativeembodiments of the invention are illustrated in the drawings, in which:

FIG. 1 illustrates a side view of a prior art sterile blender bag withliquid solution for blending with paddles to cause liquid solution toreciprocate in the general directions as shown with arrows;

FIG. 2 illustrates a top view of a prior art sterile blender bag of FIG.1 with liquid solution for blending with paddles to cause liquidsolution to reciprocate in the general directions as shown with arrows;

FIG. 3 illustrates a prior art sterile bag having regions for paddlecontact which are very close to one another to agitate fluid and anyother contents;

FIG. 4 illustrates an allograft blender bag provided with an internalscreen for retention of allograft material in a location;

FIG. 5 illustrates an allograft blender bag may be provided with aninlet port and an outlet port to handle the flow of fluid;

FIG. 6 illustrates a heat-sealable tab and screen material provided in ablender bag system;

FIGS. 7 and 8 illustrate a blender machine for selectively applyingpaddling forces to any one of the blender bags;

FIG. 9 illustrates an air purge extending through the sidewall of ablender bag;

FIGS. 10 and 11 illustrate an array of tissue treatment solutionsoperatively connected to an inlet port of the blender bag and a computercontroller for fluid delivery;

FIG. 12 illustrates an ultrasonic water bath configured to controltemperature of an allograft material; and

FIG. 13 illustrates a hollowed port system within a paddle configured toprovide temperature control.

DETAILED DESCRIPTION

Embodiments are described more fully below in sufficient detail toenable those skilled in the art to practice the system and method.However, embodiments may be implemented in many different forms andshould not be construed as being limited to the embodiments set forthherein. The following detailed description is, therefore, not to betaken in a limiting sense.

A blender and bag combination device and process is provided such thatallograft material can be effectively cleaned (e.g., removal of blood,lipid, and proteins) and disinfected (e.g., removal and killing ofbacteria and fungi). This technology can also be utilized to promotemicrobial removal of microorganisms for sampling and detection purposes.

Secondary uses of the blender and bag combination include, but are notlimited to, demineralization of bone and promotion of chondrocyteviability in joint restoration allografts.

In an embodiment, a blender device may be used to liberate microbes fromallograft material. The blender device may be a Stomacher brand devicewith modified paddles to prevent homogenization of the allograftmaterial. Traditional Stomacher brand blenders apply paddles to asterile bag (e.g., the internal surfaces of the bag are sterile) withcontents until the contents are blended into a homogenized state. Thisis an example of a destructive test that differs significantly from theblender device and methods for non-destructively testing, cleaning anddisinfecting allograft material. In either a destructive test or anon-destructive test (as described hereinbelow), Microorganisms aregenerally liberated from the tissue do to the back-and-forth directionof the liquid within the bag (i.e., the turbulent nature of the liquid.)

In one exemplary embodiment, a rectangular bag receives and retainsallograft material within the central portion of the bag using screenmaterial, which may be heat-sealed internally to the bag. The entrypoint of the allograft into the bag may also be heat sealed to create aclosed system. The bag can be filled with solution liquid (e.g.,cleaning solution, microbial extraction solution, or other suitableallograft wash solution) using bag ports. This bag system can be placedinto an additional mechanical blending system.

A motorized driven device provides paddles contact the left and rightportions of a rectangular-shaped bag. The paddles alternate back andforth at high speed to force the liquid within the bag to moveleft-to-right and right-to-left through and over the allograft materialretained in the center of the bag. This liquid turbulence removescontamination and provides excellent mechanical cleaning of theallograft material. The spent liquid can be ported off, physicallyremoving soil and contamination, and new solutions can be ported intothe bag as needed.

With reference to FIGS. 1 and 2, and as disclosed in the prior art, asterile bag 5 may be provided with liquid solution 10 for blending withpaddles 15 to cause liquid solution 10 to reciprocate in the generaldirections as shown with arrows 20. FIG. 1 illustrates a side view ofbag 5 and FIG. 2 illustrates a top view of bag 5.

With reference to FIG. 3, and as disclosed in the prior art, sterile bag5 may include regions for paddle contact 25 which are very close to oneanother to agitate fluid 10 and any other contents. This traditional bagdesign, with a traditional paddle design of a blender, causes thehomogeneous blending as referred to hereinabove. With the paddlespositioned next to one another, any soft tissue within the bag 5 willeventually be pummeled. This is true for soft tissue or any tissue. Thisis an undesirable configuration inasmuch as the allograft material willultimately be rendered unusable.

With reference now to FIG. 4, and in an embodiment, an allograft bag 30may be provided with an internal screen 35 for retention of allograftmaterial in a location 40. This positions the tissues in a centralportion of the bag and allows paddles to be separated to flankingportions beyond the position of this central portion to cause turbulentfluid flow while simultaneously avoiding tissue pummeling. This motionof the solution liquid will liberate microorganisms from the graft inthe central portion.

In an embodiment, the screen material 35 is heated-sealed into thecenter portion (location 40) allowing for the retention of allograftmaterial. This screen is designed to hold the allograft material, notallowing it to drift in the bag 30, so as to prevent paddle contact,which may crush and destroy the valuable allograft material. Likewise,the screen material 35 will allow the liquid to make contact (throughthe open pores) with the allograft material when it is forced back andforth via paddle contact with the bag. Multiple ports are integratedinto the bag whereby fluid can be added and removed from the bag asneeded.

Allograft bag 30 together with an appropriate blending machine may beused for bioburden assessment. This assessment may occur immediatelyprior to disinfection procedures to assess bioburden of the allograftmaterial. This assessment may occur immediately after disinfectionprocedures to assess bioburden of the allograft material.

With reference to FIG. 12, ultrasonic technologies may be combined withthe blender 65 depending on the type of graft. The entire system, e.g.,bag system and paddle system, may be submerged within an ultrasonicwater bath 1200. The mechanical washing from the paddles may become moreefficient when in conjunction with ultrasonic cleaning. The temperatureof the allograft material within the bag may be controlled though theultrasonic water bath. An exterior inlet port 1205 and an exterioroutlet port 1210 may be fluidly coupled with an exterior of themotorized paddle blender. Exterior inlet port 1205 and exterior outletport 1210 may be located such that the agitated fluid flows through theexterior inlet port from the ultrasonic water bath into the motorizedpaddle blender, across the outer wall of the sterile bag, and throughthe exterior outlet port back into the ultrasonic water bath.

In an embodiment, and with reference to FIG. 13, a hollowed port system1300 may be provided within the paddles 15. This port system within thepaddle may provide temperature-controlled solution 1305 constantlypumped through each of the paddles 15 to maintain a desired temperatureor temperature range during a processing embodiment. Long processingtimes with the paddling unit will most likely generate heat, thus, byporting temperature-controlled solution through the paddles 15 thecooled device may ensure constant temperature exposure (e.g., noincrease of heat) to the allograft material.

In another embodiment, a higher temperature (e.g., 45 degrees C.) may bedesired to promote higher level of cleaning and disinfection. In anotherembodiment, a lower temperature (e.g., 4 degrees C.) may be required topromote cellular viability. The hollowed port system may providetemperature regulation for either, or both, of a higher temperature or alower temperature.

In another embodiment, and with reference to FIG. 5, an allograft bag30A may be provided with an inlet port 45 and an outlet port 50 tohandle the flow of fluid 10. During operation with forces applied by oneor more paddles to one or more contact areas 25, fluid 10 may becirculated through bag 30A with a graft sealed in location 40.

Bag 30A may be loaded at the beginning of a disinfection procedure andsolution liquid may be pumped in while forces are applied by one or morepaddles to one or more contact areas 25. During each step within adisinfection procedure, bag 30A could be paddled to force thedisinfection solution in and around the graft. Because the graft isretained within the bag, disinfection procedures may require veryminimal handling of the graft.

Solution liquids of one or more types may be flushed through bag 30(e.g., not paddled) to minimize or eliminate any bacteriostatic problemsthat residual disinfection chemicals may cause for taking post-bioburdensamples.

In one embodiment, and looking at FIG. 6, a heat-sealable tab 55 andscreen material 60 may be provided in a bag 30B. Heat-sealable tab 55and screen material 60 may form the location 40 for retention ofallograft material. After placement of the allograft material within bag30B, tab 55 may be heat sealed to hold graft material. As discussedabove with respect to bag 30A, inlet port 45 and outlet port 50 may beprovided in bag 30B to handle the flow of fluid 10.

With reference now to FIGS. 7 and 8, and in an embodiment, a blendermachine 65 may be provided for selectively applying paddling forces toany one of bags 30, 30A, 30B, or 30C (discussed below.) A motorizedportion of blender machine 65 may be configured to appropriately actuateseparated paddles 15A for application of forces to the inserted bag 30,30A, 30B, or 30C. A door component 75, or other retention mechanism, maybe configured to position and maintain bag 30, 30A, 30B, or 30C withrespect to paddles 15A. As shown in FIG. 7, paddles 15A are positionedapart from one another to provide a graft retention location 40, whichallows fluid flow with adequate force and prevents the homogeneousblending of the allograft material. Bag 30, 30A, 30B, or 30C may befilled to a predetermined volume but should not be overfilled to preventbursting under paddle pressure.

In another processing embodiment, an allograft viability promotionprocess may include processing “fresh grafts” with live cells MEM(culture media)/antibiotic combinations. Fresh cartilage grafts containchondrocytes that must remain viable for successful transplantation. Analternative use of bag system with the blender system may implementminimal essential media (MEM) incubation of these fresh cartilagegrafts.

Fresh sterile MEM may be ported into the bag system to bathe the graftand promote cellular viability. The paddle system may be programmed suchthat instead of alternative paddling at high speeds to promote cleaning,a single paddle could be set to apply selective, steady force to createhydrostatic pressure (e.g., 10-100 psi) within the bag. At selectivetimes during the fresh cartilage graft incubation period, thesepressures could be applied in short intervals. This pressure wouldimitate water pressures within in vivo joints, which could promotecellular viability within the cartilage. Media changes may involve briefalternative paddling followed by fresh MEM ported into the bag after oldMEM is removed from the bag.

In one embodiment, and due to the separation of the paddles,reciprocation of the paddles may not be necessary and the paddles maypush in tandem forcing liquid from the lateral portions of bag 30, 30A,30B, or 30C to the central portion 40. In another embodiment, one of thepaddles may be configured to extend a greater distance into bag 30, 30A,30B, or 30C than the other paddle. Leaving open inlet port 45 and outletport 50, and providing an in-line stream of solution liquid 10 throughthe bag 30, 30A, 30B, or 30C. This can provide a cleaning effect and arecirculating effect (i.e., bringing in fresh solution.)

Depending on the graft material, more than one specimen could be addedto bag 30, 30A, 30B, or 30C. Additional separations could be included toeither keep fluid within an area of a particular graft or to allow fluidflow to multiple grafts (i.e., graft materials from a single donor.)

In another embodiment, and referring now to FIG. 9, an air purge 80 mayextend through the sidewall of a blender bag 30C. For initial filling ofdisinfection solution, air purge 80 is necessary to allow air to escape.Air purge 80 may include a filter to prevent contamination to the graftmaterials or solution fluid 10. Similar to the other blender bagsdescribed hereinabove, side portions for paddle contact areas 25 may beprovided by blender bag 30C. A central area 40 may be configured forretaining a graft away from paddle contact areas 25. Heat-sealable tab55 and screen material 60 may form the location 40 for retention ofallograft material in blender bag 30C. Inlet port 45 and outlet port 50may be included on bag 30C.

With reference to FIGS. 10 and 11, an array 90 of tissue treatmentsolutions may be operatively connected to inlet port 45 of bag 30C (oranother appropriate bag) and fluid delivery may be selectively operableby computer controller 95. Paddling blender machine 65 and associatedvalves may be computer controlled to maintain set process parameters.Communication conduit 100 and communication conduit 105 may be providedfrom computer controller 95 to blender machine 65 and tissue treatmentsolutions array 90, respectively. These conduits may include wired orwireless communications. A valve system 110 may be operated by computercontroller 95 to regulate fluid dispensing by array 90 and may beoperatively connected with communication conduit 100 to computercontroller 95. A communication conduit 115 may control an additionalvalve 120 between array 90 and inlet port 45. This conduit may includewired or wireless communication.

One paddle 15A may be offset with respect to the other paddle 15A so asto compress solution liquid 10 slightly more than the other paddle. Thiscauses blending and creases a pressure gradient to force solution to awaste location from outlet port 50. With the valve of outlet port 50open, and the blender 65 on, solution liquid 10 is blended acrossallograft tissue material and solution liquid 10 is slowly taken away toa waste location from outlet port 50.

With the valve of outlet port 50 closed, and the blender 65 on, solutionliquid 10 is blended across allograft tissue material and solutionliquid 10 is retained within bag 30 c instead of taken away to a wastelocation from outlet port 50.

With the valve of outlet port 50 open, and the blender 65 off, solutionliquid 10 is solution liquid 10 is run across (i.e., flushes) acrossallograft tissue material and taken away to a waste location from outletport 50 at the speed of the pull of gravity.

In an exemplary embodiment, an allograft cleaning process may includethe follow procedure. Upon sealing allograft material into the screenedportion of the bag, various detergent, disinfectant, chemical sterilant,and microbial extraction solutions may be pumped into the bag. Thissolution pumping is either stages, continuous, or a combination of bothstages and continuous solution pumping.

A stage scenario may include pumping a certain solution into the bag,executing a timed paddling process, and finally evacuating the bag. Thisprocess may be continued with additional solutions.

A continuous scenario may include pumping solutions into the bag whilethe paddling process is continuous. The evacuation port wouldessentially be left open as one or more new solutions are being pumpedinto the bag system.

If one paddle is allowed to compress further than the other paddle, itis theorized that this could create an imbalance whereby fresh solutioncould be drawn into the bag thus creating a scenario in “continuous”mode where solution is likewise forced out the outlet port. This couldmean no external pumps would be needed for this scenario.

A combination scenario may include steps that follow a staged flow andothers with a continuous flow.

An allograft microbial extraction process may include a defined speedand time of the paddles using a microbial extraction solution or sterilewater. High extraction efficiency may be achieved using a paddlingmethodology. The solution taken from the outlet port may be used formicrobial detection analysis (e.g., real time PCR, fluorescent laserscanning, traditional microbiological analysis, etc.)

In another processing embodiment, an allograft demineralization processmay be provided in addition to cleaning allograft material. Depending onthe solution fluid, e.g., an acid solution, the mechanical washingeffect of the bag paddling could also provide efficient bonedemineralization. The paddle agitation surrounding the mineralizedallograft material may be advantageous as fresh solution is ported intothe bag and allowing for “spent” acid solution to be ported away.

Additionally, all spent solutions ported off from the bag system in maybe taken for microbiological analysis to verify in process control ofthe allograft cleaning process.

In another embodiment, an allograft final packaging process may beprovided using the bag system. One of the advantages of the bag systemis containing the tissue in a closed environment. Following apost-processing final tissue assessment, which may involve technicianhandling and any final debridement, the tissue may be placed into a newsterile bag system and then undergo some final disinfection and rinsingsteps. The bag system could be then be minimally modified (e.g., portssealed, etc.) to become the final packaging for the allograft material.

Paddling blender 65 and any of bags 30, 30A, 30B, or 30C may be sizedlarge enough to disinfect all currently processed graft sizes.

Paddling Parameters

To obtain adequate fluid motion within the bag to initiate microorganismrelease from the tissue graft or the tissue grafts, in an embodiment,the paddling may be between 300-800 paddle compressions a minute forboth of the paddles. Using a stomacher machine, 1 rotation provides 2compressions (i.e., one compression for each paddle per rotation.) Thisconfiguration may provide 150 to 600 paddle compressions for a stomachermachine with an operating range of 75-300 RPM. Other operating rangesmay provide other amounts of paddle compressions. The shear forcesgenerated within this range will be sufficient to remove microorganismcontamination from the tissue grafts(s).

Ultrasonic Parameters

If this device is used within an ultrasonic water bath, the unit must beoutputting at a frequency of 40-150 kHz at a power setting of 50-300Watts per gallon.

Flow Parameters

This setting can be highly variable depending on the desire addition andremoval of solution from the bag. Optimally, a setting of 1 mL persecond will suffice.

Temperature Parameters

The solution within the bag system may be operated between 4-55 degreesC. depending on the desired temperature setting. Higher temperatures aredesired for microorganism kill and lower temperature are desired forallograft cell viability (e.g., chondrocyte, fibroblasts).

Example 1

The following processing example relates to processing a fresh cartilagegraft. This is a processing example, thus, types of solutions, steps,and processing times may be changed or modified.

Tissue may undergo typical debridement and processing steps wherebysurrounding unwanted tissue are removed from the cartilage. Any otherprocessing could occur at this stage (e.g., centrifugation).

Debrided grafts may be placed into the bag system and the bag may besealed.

MEM solution may be ported into the bag using a staged processingscenario. The paddles may be maintained at a temperature of 22 degreesC. using flowing solution. For 5 minutes, the paddles are moved at highspeed. An exemplary range of high-speed paddle movement is 300 RPM. Thesolution may be ported off (and may be sampled for microbialcontamination.)

MEM solution containing an antibiotic cocktail may be ported into thebag system. The flow juncture to the paddles may be switched to maintaina temperature of 32 degrees C. For 5 minutes, the paddle will be allowedto move at high speed. The solution may sit unanimated for the following15 minutes. This may be repeated (5 minute paddling+15 incubation) twomore times to make a total MEM antibiotic exposure 1 hour. The solutionmay be ported off (and may be sampled for microbial contamination).

MEM solution containing an antibiotic cocktail may be ported into thebag system. The flow juncture to the paddles may be switched to maintaina temperature of 32 degrees C. For 5 minutes, the paddles will beallowed to move at high speed. The solution may sit unanimated for 15minutes. This may be repeated (5 minute paddling+15 incubation) two moretimes to make a total MEM antibiotic exposure 1 hour. The solution maybe ported off (and may be sampled for microbial contamination).

MEM solution (no antibiotics) may be ported into the bag system. Theflow juncture of the paddles may be switched to maintain a temperatureof 22 degrees C. No paddling occurs for a 5-minute exposure. Thesolution may be ported off (and may be sampled for microbialcontamination).

The bag would be opened to allow for the technician to perform any finaldebridement and/or “other/outside” of the bag system processing stepswith the tissue. The bag will be discarded.

The graft may be placed into a fresh sterile bag system. MEM solution(no antibiotics) may be ported into the bag system. A two-minuteexposure to the high-speed paddles may be followed by the MEM solutionported off into a sterile vessel for microbial contamination assessment.This may be designated as the final/pre-incubation culture.

MEM solution containing Bacitracin (50 U/mL) and Polymyxin B (500 U/mL)may be ported into bag system. The flow juncture to the paddles may beswitched to maintain a temperature of 4 degrees C. AlloPulse may be setto perform incubation processing described in Process Embodiment D. Oncean hour, the bag may be gently compressed to achieve a positivehydrostatic pressure within the bag system solution of 25 psi (this holdwill last two minutes). The solution may be held in the bag for no morethan 48 hours.

On or before 48 hours (4 degrees C.), the MEM will be ported off(solution can be assayed for contamination-real time analysis) from thebag system and fresh MEM containing Bacitracin/Polymyxin B may be portedinto the bag. Once an hour, the bag may be gently compressed to achievea positive hydrostatic pressure within the bag system of 25 psi (thishold will last two minutes). The solution will be held in the bag for nomore than 48 hours.

A final real-time microbial assessment can be made immediately prior tograft release.

With the bag system contain the graft and MEM with Bacitracin/PolymyxinB, the bag system may be sealed off and become the final packaging usedto distribute the graft.

Example 2

The following processing example is a reflection of above-mentionedembodiments with regard to processing a traditional allograft (e.g.,tendon or tricortical wedge). This is merely a processing example, thustypes of solutions, steps, and processing times may change.

Tissue may undergo typical debridement and processing steps wherebysurrounding unwanted tissues are removed from the tissue. Shaping andcutting of allograft may be performed. Any other pre-bag systemprocessing could occur at this stage (e.g., centrifugation). It isrecommended that a centrifugation step precede the bag system steps.

Debrided and shaped grafts would be placed into the bag system and thebag may be sealed.

Bacitracin (50 U/mL)/Polymyxin B (500 U/mL) cocktail solution may beported into the bag using a staged processing scenario. The paddles maybe maintained at a temperature of 37 degrees C. using flowing solution.For 5 minutes, the paddles are moved at high speed. In an exemplaryembodiment, the range of high-speed paddle movement may be 300 RPM. Thesolution may sit unanimated for 15 minutes. This may be repeated (5minute paddling+15 minute incubation) two more times to make totalBacitracin/Polymyxin B cocktail exposure 1 hour. The solution may beported off (and may be sampled for microbial contamination.) This stepmay be repeated if necessary.

3% Hydrogen Peroxide solution may be ported into the bag using a stagedprocessing scenario. The paddles may be maintained at a temperature of45 degrees C. using flowing solution. For 5 minutes, the paddlesoperated at high speed. The solution may sit unanimated for 15 minutes.This may be repeated (5 minute paddling+15 incubation) two more times tomake total 3% H₂O₂ cocktail exposure for 1 hour. In an embodiment, aone-way check valve may be provided through the bag for outgassing. Anadditional tube may be attached to inlet port 45 and may be positionedabove the bag to allow only gas to vent from the bag. The solution maybe ported off (and could be sampled for microbial contamination.) Thisstep may be repeated if necessary.

The bag may be opened to allow for the technician to perform any finaldebridement and/or “other/outside” processing steps with the tissue. Thebag may be discarded.

The graft may be place into a fresh sterile bag system. Sterile watermay be ported into the bag system. For 5 minutes, the paddles areoperated at high speed. The solution may sit unanimated for 15 minutesfollowed by 5-minute high-speed paddle step. The solution may be portedoff (and could be sampled for microbial contamination). This step may berepeated if necessary.

Sterile water may be ported into the bag system. A two-minute exposureto the high-speed paddles may be followed by the water ported off into asterile vessel for microbial contamination assessment. This may bedesignated as the final culture.

The bag system containing the graft may be sealed off, frozen down, andbecome the final packaging used to distribute the graft.

Example 3

The following processing example is a reflection of above-mentionedembodiments with regard to demineralizing bone. This is merely aprocessing example, thus, types of solutions, steps, and processingtimes may change.

Milled tissues to particular specifications (e.g., particle range) maybe added into the bag system and the bag may be sealed. The pore sizewithin the screened material within the center of the bag may need to beof sufficient size not to let said particle range escape from the centerportion of the bag.

0.5 N Hydrochloric Acid would be ported into the bag using a stagedprocessing scenario. The paddles would be maintained at a temperature of22 degrees C. using flowing solution. For 5 minutes, the paddles mayoperated at high speed. The solution may sit unanimated for 15 minutesfollowed by a 5-minute high-speed paddle step. The solution may beported off (and may be sampled for residual calcium), This step may berepeated as necessary.

Sterile Water may be ported into the bag using a staged processingscenario. The paddles may be maintained at a temperature of 22 degreesC. using flowing solution. For 5 minutes, the paddles will be operatedat high speed. The solution may sit unanimated for 15 minutes followedby a 5-minute high-speed paddle step. The solution may be ported off(and may be sampled for residual calcium.) This step may be repeated asnecessary.

Sodium Phosphate Buffer Solution may be ported into the bag using astaged processing scenario. The paddles may be maintained at atemperature of 22 degrees C. using flowing solution. For 5 minutes, thepaddles may be operated at high speed. The solution may sit unanimatedfor 15 minutes followed by a 5-minute high-speed paddle step. Thesolution may be ported off (and may be sampled for residual calcium).This step could be repeated as necessary.

Sterile Water may be ported into the bag using a staged processingscenario. The paddles may be maintained at a temperature of 22 degreesC. using flowing solution. For 5 minutes, the paddles may be operated athigh speed. The solution would sit unanimated for 15 minutes followed bya 5-minute high-speed paddle step. The solution would be ported off (andcould be sampled for residual calcium). This step could be repeated asnecessary.

Since demineralization products are usually freeze-dried, the tissuewould be removed from the bag system for freeze-drying process orprocesses. The freeze-drying process is not a part of the bagsystem/blender system technology.

Although the above embodiments have been described in language that isspecific to certain structures, elements, compositions, andmethodological steps, it is to be understood that the technology definedin the appended claims is not necessarily limited to the specificstructures, elements, compositions and/or steps described. Rather, thespecific aspects and steps are described as forms of implementing theclaimed technology. Since many embodiments of the technology can bepracticed without departing from the spirit and scope of the invention,the invention resides in the claims hereinafter appended.

What is claimed is:
 1. A system for cleaning and disinfecting allograftmaterial, the system comprising: a sterile bag having an outer wallconfigured to withstand compressive forces, a pair of paddle contactsections, and an allograft retaining section separate from the paddlecontact sections; a motorized paddle blender having at least twopaddles, each having a compressive surface configured to applycompressive forces onto the sterile bag at the pair of paddle contactsections, a motorized portion to actuate the at least two paddles, and adoor component configured to hold the sterile bag adjacent the at leasttwo paddles, wherein the at least two paddles and the allograftretaining section are configured to prevent the at least two paddlesfrom compressing the allograft material when the compressive forces areapplied onto the sterile bag at the pair of paddle contact sections; anda hollowed port system within each of the at least two paddles, thehollowed port system comprising inflow and outflow ports in fluidcommunication with a hollow interior of each of the at least twopaddles, wherein throughout an actuation cycle of the at least twopaddles, temperature-controlled solution constantly flows into each ofthe inflow ports, across the compressive surfaces of each of the atleast two paddles, and out each of the outflow ports, providing aconstant flow of the temperature-controlled solution across thecompressive surfaces of each of the at least two paddles such that thecompressive surfaces resist frictional heating and maintain a constanttemperature throughout the actuation cycle; and further comprising aninternal screen defining the allograft retaining section at a centrallocation between the pair of paddle contact sections, the centrallocation defined by the internal screen configured to retain theallograft tissue between the pair of paddle contact sections to preventtissue pummeling and to allow turbulent fluid flow therethrough, whereinthe door component, the at least two paddles, the pair of paddle contactsections, and the central location of the allograft retaining sectionare configured with respect to one another to locate the compressiveforces onto the sterile bag on the pair of paddle contact sections atpositions away from the central location of the allograft retainingsection so as to in turn provide a non-contact area at the centrallocation to prevent application of the compression forces to theallograft material.
 2. A system in accordance with claim 1, wherein thesterile bag is a paddle blender bag having modifications to provide theallograft retaining section separate from the pair of paddle contactsections.
 3. A system in accordance with claim 1, wherein the at leasttwo paddles are separated by a given distance to apply the compressiveforces on the sterile bag at the pair of paddle contact sections, andwherein the allograft retaining section has a width less than the givendistance separating the at least two paddles so as to allow theallograft retaining section to retain the allograft material between thepair of paddle contact sections corresponding to the at least twopaddles.
 4. A system for cleaning and disinfecting allograft material,the system comprising: an ultrasonic water bath containing an amount ofagitated fluid to receive and submerge a sterile bag mounted within amotorized paddle blender, wherein: the sterile bag has an outer wallconfigured to withstand blending forces, at least one paddle contactsection, and an allograft retaining section separate from the at leastone paddle contact section; and the motorized paddle blender has atleast one paddle configured to apply blending forces onto the sterilebag at the at least one paddle contact section, a motorized portion toactuate the at least one paddle, a door component configured to hold thesterile bag adjacent the at least one paddle, and an exterior inlet portand an exterior outlet port fluidly coupled with an exterior of themotorized paddle blender and located such that the agitated fluid flowsthrough the exterior inlet port from the ultrasonic water bath into themotorized paddle blender, across the outer wall of the sterile bag, andthrough the exterior outlet port back into the ultrasonic water bath,wherein the at least one paddle and the allograft retaining section areconfigured to prevent the paddle from contacting the allograft materialwhen the blending forces are applied onto the sterile bag at the atleast one paddle contact section.
 5. A system in accordance with claim4, wherein the ultrasonic water bath is configured to controltemperature of the allograft material.
 6. A system in accordance withclaim 5, wherein the ultrasonic water bath is configured to transmitultrasonic vibrations through the outer wall of the sterile bag tosupplement mechanical washing of the allograft material with forces offluid created by the at least one paddle.
 7. A system in accordance withclaim 4, further comprising an internal screen defining the allograftretaining section at a location adjacent to the at least one paddlecontact section, the location defined by the internal screen configuredto retain the allograft tissue adjacent to the paddle contact section toprevent tissue pummeling and to allow turbulent fluid flow therethrough,wherein the door component, the at least one paddle, the paddle contactsection, and the location of the allograft retaining section areconfigured with respect to one another to locate the blending forcesonto the sterile bag on the paddle contact section at a position awayfrom the allograft retaining section so as to in turn provide anon-contact area at the allograft retaining section to preventapplication of the blending forces to the allograft material.